Custom antibody development

We offer world-leading antibody development platforms based on a proprietary RabMAb® rabbit monoclonal platform, a phase display platform (AxioMx), and a next-generation sequencing platform (NGS-RabMAb®)

Applications

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application

Abreviews

Notes

Dot blot

1/1000.

WB

1/5000 - 1/20000. Predicted molecular weight: 11 kDa.

ICC/IF

1/500 - 1/2000.

ChIP

Use at an assay dependent concentration. 2 - 8 ul per IP

IP

Use at an assay dependent concentration.

Target

Function

Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.

Sequence similarities

Belongs to the histone H4 family.

Post-translationalmodifications

Acetylation at Lys-6 (H4K5ac), Lys-9 (H4K8ac), Lys-13 (H4K12ac) and Lys-17 (H4K16ac) occurs in coding regions of the genome but not in heterochromatin.Citrullination at Arg-4 (H4R3ci) by PADI4 impairs methylation.Monomethylation and asymmetric dimethylation at Arg-4 (H4R3me1 and H4R3me2a, respectively) by PRMT1 favors acetylation at Lys-9 (H4K8ac) and Lys-13 (H4K12ac). Demethylation is performed by JMJD6. Symmetric dimethylation on Arg-4 (H4R3me2s) by the PRDM1/PRMT5 complex may play a crucial role in the germ-cell lineage.Monomethylated, dimethylated or trimethylated at Lys-21 (H4K20me1, H4K20me2, H4K20me3). Monomethylation is performed by SET8. Trimethylation is performed by SUV420H1 and SUV420H2 and induces gene silencing.Ubiquitinated by the CUL4-DDB-RBX1 complex in response to ultraviolet irradiation. This may weaken the interaction between histones and DNA and facilitate DNA accessibility to repair proteins. Monoubiquitinated at Lys-92 of histone H4 (H4K91ub1) in response to DNA damage. The exact role of H4K91ub1 in DNA damage response is still unclear but it may function as a licensing signal for additional histone H4 post-translational modifications such as H4 Lys-21 methylation (H4K20me).Sumoylated, which is associated with transcriptional repression.

Anti-Histone H4 (acetyl K12) antibody - ChIP Grade images

ChIP - Anti-Histone H4 (acetyl K12) antibody (ab46983)

ab46983 tested by ChIP analysis. ChIP performed using Hela chromatin (1.5 x 106 cell equivalents per ChIP) and 10 ul of ab46983 or the equivalent amount of rabbit IgG as a negative control. Real time, quantitative PCR (RT-qPCR) was performed on DNA purified from each of the ChIP reactions using a primer pair specific for the indicated gene. Data are presented as fold enrichment of the ChIP antibody signal versus the negative control IgG using the ddCT method.

ab46983 in dot blot (top panel), acetylated peptides corresponding to the immunogen and related peptides were spotted onto PVDF and probed at a dilution of 1:1000. The amount of peptide (picomoles) spotted is shown next to each row.